Objective To analyse the cloning and sequencing of mature fragment of human bone morphogenetic protein-4 gene. Methods The template DNA was extract from the human osteosarcoma cells line U-2OS by the single-step isolation method with isothiocyanic acid guanidine, the cDNA coding for the mature fragment of BMP-4 was amplified by the reverse transcription-polymerase chain reaction(RT-PCR). The mature fragment of BMP-4 was cloned into the vector pUC19, and sequenced by Sanger dideoxy-mediated chain termination method. Results The mature fragment of BMP-4 cDNA was obtained by RT-PCR and identified by sequencing. The computer search was done on Genebank, and the published DNA sequence of BMP-4 from Genebank (D30751) was chosen for a reference. Analysis showed that the homology and similarity of nucleotides and amino acids between cDNA of rhBMP-4 mature fragment and the published sequence of BMP-4 were both 99％ . Sequence analysis revealed that there were two bases mutations, one was at base 1 154(201) G C, this had no influence on the corresponding amino acids(Val), another was at base 1 222(269) C T, the mutation at the base 1 222 had turned the Ala into Val. Conclusion The mature fragment of BMP-4 gene has been cloned. The gene is of great significance in treatment of skeletal injuries and diseases.

ObjectiveTo study the cloning and sequencing of mature fragment of human bone morphogenetic protein-4 gene. Methods The template DNA was obtained from the human osteosarcoma cell line U2OS. By using RT- PCR method, the cDNA coding for the mature fragment of BMP-4 was amplified, cloned into the vector pUC19, and sequenced by Sanger Dideoxy-mediated Chain Termination method. Results The mature fragment of BMP4 cDNA was obtained by RT-PCR and determined by sequencing. Through the computer search on Genebank, the analysis showed that the homology of nucleotides and amino acids between cDNA of rhBMP4 mature fragment of this study and the published sequence was 99％. Sequence analysis showed that there were two differences, one was at base 1154 (201): G→C, which had no influence on the corresponding amino acids (Val). Another was at basel222 (269):C→T, the mutation at the base 1222 had the change of Ala to Val. Conclusion The mature fragment of BMP4 gene has been cloned. The results will be of great significance in treatment of skeletal injuries and diseases.

ObjectiveTo compare the effect of two analgesia methods in silver needle therapy among patients with myofascitis.Methods Totally 181 eligible patients with myofascitis were collected from January 2015 to March 2016 and were divided into observation group and control group by random number table. Patients in the control group (n=90) were given partial external lidocaine ointment and diclofenac sodium lidocaine hydrochloride intramuscular injection an hour before the treatment. Patients in the observation group (n=91) first received partial external lidocaine ointment an hour before the treatment,then received flurbiprofen axetil intravenous infusion and intravenous drip 15 minutes before the treatment. Visual analogue scale (VAS) was applied to evaluate the pain that before treatment,at the moment of acupuncture,during the treatment,at the moment of pulling out the needles,and a day after the treatment. Moreover,the clinical effect of two groups was also evaluated after a course of treatment.Results The VAS scores at the moment of acupuncture and during the treatment of the observation group were (7.71±0.80) and (6.71±1.17),while the data in the control group were (4.14±1.06) and (3.58±0.84) (P<0.05). There were no significant differences in the VAS scores before treatment,after pulling out the needles,and a day after treatment,and the clinical effect between two groups (P>0.05).Conclusions Flurbiprofen axetil injection can effectively reduce the pain in silver needle therapy among patients with myofascitis and can also improve patients′ compliance.